Effects of sphingomyelin and cholesterol on lipoprotein lipase-mediated lipolysis in lipid emulsions.
Lipoprotein lipase (LPL) hydrolyzes triacylglycerol (TG) of TG-rich lipoproteins. We investigated the effects of sphingomyelin (SM) and cholesterol (Chol) on the lipolysis of lipid emulsions by LPL using human apolipoprotein C-II (apoC-II) or plasma as an activator. Kinetic studies of the lipolysis rates clearly demonstrated that the primary effect of the activator on the LPL reaction was not to increase the affinity of LPL for the emulsion surface, but to enhance LPL catalytic activity. Incorporation of SM into the emulsion surface caused increases in Km(app) and decreases in Vmax(app), indicating that SM inhibited lipolysis by decreasing both affinity for substrates and catalytic activity of LPL. SM was also found to affect possible factors related to the lipolysis rates; that is, SM increased TG solubility in surface layers and decreased apoC-II binding to the emulsion surface. Interestingly, Chol did not affect the lipolysis rates even though it decreased TG solubility and apoC-II binding. These results indicated that neither TG solubility nor amount of apoC-II binding were determinate factors in LPL-mediated lipolysis under physiological conditions. Our results suggest that the content of SM in the lipoprotein surface plays an important role by controlling lipoprotein lipase-mediated lipolysis, and that cholesterol enrichment in the lipoprotein surface has no influence on lipolysis, but may affect other metabolic processes such as uptake by the liver through the selectivity of apolipoprotein binding.
Arimoto
I
I
Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Saito
H
H
Kawashima
Y
Y
Miyajima
K
K
Handa
T
T
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C / Q000378:metabolism / Q000494:pharmacology; D001769:Blood; D002784:Cholesterol / Q000494:pharmacology*; D004655:Emulsions; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D050356:Lipid Metabolism; D008066:Lipolysis / Q000187:drug effects*; D008071:Lipoprotein Lipase / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D012995:Solubility; D013109:Sphingomyelins / Q000494:pharmacology*; D014280:Triglycerides / Q000378:metabolism; D014304:Triolein / Q000378:metabolism
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C; D004655:Emulsions; D013109:Sphingomyelins; D014280:Triglycerides; D014304:Triolein; D002784:Cholesterol; D008071:Lipoprotein Lipase